Large-scale isolation of proteins of the large subunit from Escherichia coli ribosomes.

A strategy has been developed and optimized that allows the isolation of proteins of the large subunit from Escherichia coli ribosomes and combines the following advantages: speed, applicability for the isolation of milligram amounts of a single protein, and preservation of the biological activity of the proteins. The method consists of the following steps: ion-exchange chromatography on MonoS and MonoQ, gel filtration on Sephadex 75, and salt washes. Eleven proteins can be purified by a single chromatographic step, and a combination of two steps enables the isolation of the other proteins.

[1]  J. Frank,et al.  A model of protein synthesis based on cryo-electron microscopy of the E. coli ribosome , 1995, Nature.

[2]  M van Heel,et al.  The 70S Escherichia coli ribosome at 23 A resolution: fitting the ribosomal RNA. , 1995, Structure.

[3]  D. Svergun,et al.  Solution scattering from 50S ribosomal subunit resolves inconsistency between electron microscopic models. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R P May,et al.  Inter‐protein distances within the large subunit from Escherichia coli ribosomes. , 1992, The EMBO journal.

[5]  K. Nierhaus,et al.  The assembly of prokaryotic ribosomes. , 1991, Biochimie.

[6]  W. S. Champney.  Reversed-phase chromatography of Escherichia coli ribosomal proteins. Correlation of retention time with chain length and hydrophobicity. , 1990, Journal of chromatography.

[7]  K. Nierhaus.  The allosteric three-site model for the ribosomal elongation cycle: features and future. , 1990, Biochemistry.

[8]  Harry F. Noller,et al.  Intermediate states in the movement of transfer RNA in the ribosome , 1989, Nature.

[9]  R. Brimacombe,et al.  A model for the spatial arrangement of the proteins in the large subunit of the Escherichia coli ribosome. , 1988, The EMBO journal.

[10]  J A Langer,et al.  A complete mapping of the proteins in the small ribosomal subunit of Escherichia coli. , 1987, Science.

[11]  K. Nierhaus,et al.  Incorporation of six additional proteins to complete the assembly map of the 50 S subunit from Escherichia coli ribosomes. , 1987, The Journal of biological chemistry.

[12]  K. Nierhaus,et al.  50-S subunit from Escherichia coli ribosomes. Isolation of active ribosomal proteins and protein complexes. , 1979, European journal of biochemistry.

[13]  A. Liljas,et al.  Small‐angle X‐ray scattering and crosslinking study of the proteins L7/L12 from Escherichia coli ribosomes , 1976, FEBS letters.

[14]  T. Nakamoto,et al.  Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate. , 1972, The Journal of biological chemistry.

[15]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[16]  M. Garber,et al.  Proteins of the Thermus thermophilus ribosome. Purification of proteins from the large ribosomal subunit. , 1994, Biochimie.

[17]  G. Spedding.  Ribosomes and protein synthesis : a practical approach , 1990 .

[18]  H. Rheinberger,et al.  Parameters for the preparation of Escherichia coli ribosomes and ribosomal subunits active in tRNA binding. , 1988, Methods in enzymology.

[19]  B. Cooperman,et al.  Reversed-phase high-performance liquid chromatography of ribosomal proteins. , 1988, Methods in enzymology.

[20]  P. Nowotny,et al.  Preparation and activity measurements of deuterated 50S subunits for neutron-scattering analysis. , 1988, Methods in enzymology.

[21]  M. Capel,et al.  Ion-exchange high-performance liquid chromatographic separation of ribosomal proteins. , 1988, Methods in enzymology.